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Soluble Human M-CSF Receptor and Uses Thereof

a human mcsf and receptor technology, applied in the field of soluble human mcsf receptors, can solve the problems of significant increase in cancer morbidity and mortality

Inactive Publication Date: 2010-04-22
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016]In other embodiments of the invention, the soluble M-CSF receptor levels detect the presence of, predict prognosis or severity of, or monitor the progression of a disease selected from the group consisting of osteolytic diseases associated with relatively increased osteoclast activity or the osteoclastic/osteoblastic process required for bone remodeling, including metabolic bone diseases, endocrinopathies (hypercortisolism, hypogonadism, primary or secondary hyperparathyroidism, hyperthyroidism), hypercalcemia, deficiency states (rickets/osteomalacia, scurvy, malnutrition), ch

Problems solved by technology

In general, cancer morbidity and mortality increases significantly if it is not detected early in its progression.

Method used

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  • Soluble Human M-CSF Receptor and Uses Thereof
  • Soluble Human M-CSF Receptor and Uses Thereof
  • Soluble Human M-CSF Receptor and Uses Thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0093]This example describes the identification of a soluble receptor for human M-CSF and shows that this soluble receptor is capable of binding M-CSF.

[0094]I. Identification of Soluble Human M-CSF Receptor in Serum Samples

[0095]A. Materials and Methods

[0096]Micro titer plates (R&D systems, Cat# CP0011) were coated with 100 μl / well capture antibody (Duo Set Elisa development system hM-CSF R(R&D systems, Cat# DY329)) at a working dilution of 100 ng / ml in PBS. The plates were sealed and incubated at room temperature overnight. After washing the wells 3 times with wash buffer (0.05% Tween in PBS, pH 7.2-7.4) with a manifold dispenser / washer, plates were blocked with 300 μl / well of blocking buffer (1% BSA, 5% Sucrose in PBS, pH 7.2-7.4) at room temperature for at least 1 hour.

[0097]After repeating the wash step, the plates were ready for sample addition. 100 μl / well of either sample or standards in reagent diluent were added. Plates were covered with adhesive and incubated for 2 hours a...

example 2

[0115]This Example shows that the soluble M-CSF receptor was found in human urine samples. This Example further shows that this receptor is present in serum from both normal subjects and breast cancer patients, and the soluble M-CSF receptor level correlated with the M-CSF level. Finally, this Example shows that the soluble M-CSF receptor is also found in primates.

[0116]I. Soluble Human M-CSF Receptor was also Found in Urine Samples

[0117]Using the same ELISA setup as described in Example 1, multiple urine samples from healthy human volunteers were collected and assayed for the presence of soluble human M-CSF receptor. Varying levels of soluble human M-CSF receptor were detected among adult female (age from 25 to 66), adult male (age from 35 to 70) and children from age 3 to 9. Quantification of the concentration of soluble human M-CSF receptor was determined with recombinant c-fms-human Fc fusion protein as reference (FIG. 9).

[0118]Urine samples were collected daily throughout the m...

example 3

[0125]This Example shows that the expression of soluble human M-CSF receptor is tightly coupled with osteoclast differentiation. This Example further shows that only differentiated osteoclasts express the soluble human M-CSF receptor, the level of which increases upon removal of M-CSF.

[0126]A. Materials and Methods

[0127]The expression of M-CSF soluble receptor was studied in an in vitro system with human osteoclast cells, where the expression of the membrane-bound M-CSF receptor was found. In particular, the expression of the soluble receptor was examined in the context of osteoclast differentiation. Primary human Osteoclast Precursors (Cambrex Bio Science Walkersville, Inc. Cat# 2T-110) were seeded at 10,000 cells / well at 0.2 ml / well in the cell culture media containing 30 ng / ml human M-CSF and 100 ng / ml RANKL. Test antibodies were added to the wells at 1 μg / mL on the same day of cell plating. The cells were incubated at 37° C., in a humidified atmosphere of 5% CO2. On day 7, osteo...

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Abstract

Soluble human M-CSF receptor is provided, along with pharmaceutical compositions containing such receptor, kits containing a pharmaceutical composition, and methods of diagnosing and treating diseases and disorders associated with M-CSF such as bone loss in a subject afflicted with an osteolytic disease.

Description

TECHNICAL FIELD[0001]This invention relates to a naturally occurring soluble fragment of the receptor for human M-CSF which is capable of binding M-CSF, methods of using this soluble fragment for therapy, and methods of diagnosing diseases and conditions by detecting this soluble fragment in patient samples.BACKGROUND OF THE INVENTION[0002]An estimated 1.4 million new cases of cancer occur every year, and an estimated 0.6 million people die from cancer every year. With improvements in detection and treatment, many of these patients survive for significant periods of time. As of Jan. 1, 2002, there were an estimated 10.1 million cancer survivors, representing approximately 3.4% of the population. Of these cancer survivors, breast (22%), prostate (18%), colorectal (10%) and gynecologic (10%) are the most common cancer sites.[0003]In general, cancer morbidity and mortality increases significantly if it is not detected early in its progression. Treatment decisions are often linked to th...

Claims

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Application Information

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IPC IPC(8): G01N33/53
CPCG01N33/574G01N33/57488G01N33/6893G01N2333/535G01N33/689G01N33/50G01N33/48G01N33/487
Inventor LIU, CHENGDEUTER-REINHARD, MAJAKUNICH, JOHNKAVANAUGH, MICHAEL
Owner NOVARTIS AG
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